Studies on the penetration of mammalian cells by deoxyribonucleoside‐5′‐phosphates

Waqar, M. Anwar; Taber, Robert; Huberman, Joel A.

doi:10.1002/jcp.1041010207

Cited by 6 publications

(5 citation statements)

References 23 publications

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“…Permeation of intact exogenous nucleotides through the plasma membrane of mammalian cells does not normally occur in an easily detectable fashion. Nevertheless, mammalian cells can be infected with low efficiency by viral nucleic acids (Holland et al, 19591, and mouse fibroblasts (L cells) were shown to be permeable to intact dAMP and araAMP (Plunkett and Cohen, 19771, as well as other deoxynucleoside monophosphates (Waqar et al, 1979). Incorporation of low levels of intact, exogenously supplied nucleotides into the cellular pools is impaired by the activity of ectoenzymes such as ATPase, ADPase, alkaline phosphatase, and 5' -nucleotidase (Stanley et al, 1982).…”

Section: Discussionmentioning

confidence: 99%

Treatment of human tumor cells with ADP or ATP yields arrest of growth in the S phase of the cell cycle

Rapaport

1983

Journal Cellular Physiology

126

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Treatment of a variety of cultured human tumor cells with low levels (40-80 pM) of adenosine 5' -diphosphate (ADP) or adenosine 5' -triphosphate (ATP) has produced arrest of these cells in the S phase of their cycle followed by cellular death. ADP and ATP are demonstrated to be incorporated into the cellular acid-soluble nucleotide pools, presumably by permeation through the plasma membrane of these cells. Since AMP, adenosine, 3 ' , 5' cyclic AMP, or a variety of diadenosine polyphosphates do not produce similar cytostatic effects or similar alterations of the cellular acid-soluble nucleotide pools, we suggest that the effects of ADP and ATP are not due to their prior breakdown or modification. Cultured animal cells have been widely acknowledged not to incorporate acid-soluble adenine nucleotides although incorporation of exogenous intact nucleoside monophosphate into cellular pools has been demonstrated in a few cases. A 48-hour treatment of logarithmically growing h u m a n tumor cells with 40 pM of ADP or ATP produced substantial arrest of cell populations in the S phase of their cycle and a dramatic reduction in total cellular uridine 5' -triphosphate (UTP) pools. AMP, adenosine, 3',5' cyclic AMP, Ap3A, Ap4A or Ap5A produced small and dissimilar effects. The experiments reported here suggest that the plasma membranes of several lines of human tumor cells are permeable to low levels of intact ADP and ATP. The suggestion of en bloc incorporation of low ADP and ATP levels into human tumor cells is supported by the almost identical effects of ADP and ATP on total cellular acid-soluble nucleotide pools and cellular growth. The rapid conversion of intracellular ADP pools into ATP in tumor cells has been reported.

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Section: Discussionmentioning

confidence: 99%

Treatment of human tumor cells with ADP or ATP yields arrest of growth in the S phase of the cell cycle

Rapaport

1983

Journal Cellular Physiology

126

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“…Even though reverse transcription is more efficient in the presence of detergents, significant quantities of intravirion DNA can be synthesized without detergent and can subsequently augment viral infectivity. It has been suggested that the cellular membrane may not be significantly permeable to either ribonucleoside triphosphates or dNTPs (19,25,29,32). The retroviral envelope is derived from the cellular membrane of virus-producing cells.…”

Section: Figmentioning

confidence: 99%

Increasing transduction efficiency of recombinant murine retrovirus vectors by initiation of endogenous reverse transcription: potential utility for genetic therapies

Zhang

Duan

Dornadula

et al. 1995

J Virol

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Reverse transcription of retroviral genomic RNA in a target cell is influenced by cellular factors, including the concentration of deoxyribonucleoside triphosphates (dNTPs). In addition, recent data have demonstrated that reverse transcription can be driven within human immunodeficiency virus type 1 virions, prior to infection of a cell, by increasing extracellular concentrations of dNTPs. In attempts to increase the transduction efficiency of recombinant murine leukemia virus vectors, endogenous reverse transcription was initiated within cell-free, recombinant murine leukemia virus virions in the presence of relatively high concentrations of dNTPs. As a result, the expression of transduced genes via these retroviral vectors was increased approximately 10-fold by treatment of virions with dNTPs. Combined with our previous data, these observations suggest that virion-associated DNA synthesis can occur in diverse groups of retroviruses and positively alter retroviral infectivity. As such, these manipulations may be useful for increasing the efficiency of retrovirus-mediated gene delivery.

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“…28). The rate-limiting step governing uridine incorporation is likely to be at the level of uridine kinase (29).…”

Section: Enhanced Incorporation Of [3h]itridine As a Measitre O F Celmentioning

confidence: 93%

Lectin pulses as determinants of lymphocyte activation and inactivation during the first six hours of culture: sequential action of concanavalin A and complement cause cell lysis

Forsdyke¹

1980

Can. J. Biochem.

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Rat lymph node cells were incubated for 6 h in medium containing bovine calf serum. Cell activation or inactivation by concanavalin A (Con A) was assessed by measuring changes in radioactive labelling with [3H]uridine. Two inactivation processes were distinguished: complement-dependent inactivation needing moderately high Con A concentrations (500 μg/mL serum) and complement-independent inactivation needing very high Con A concentrations. The times needed for cells to react with sufficient Con A to produce activation and the complement-dependent inactivation were compared by adding extra serum at different times to bind Con A and reduce the effective Con A concentration. Cells exposed to Con A prior to adding serum rapidly reacted with sufficient Con A to produce optimum activation and inactivation. Cells exposed to Con A in the presence of serum reacted with Con A at a lower rate. Initial reaction of cells with Con A was dependent on time and Con A concentration in a reciprocal manner (i.e., a high concentration of Con A could compensate for a short pulse time). Inactivation of cells was dependent on the concentrations of Con A and serum (complement) in a reciprocal manner (i.e., a high concentration of Con A compensated for a low concentration of serum). Studies with complement inhibitors added at different times of culture showed that, whereas cell activation begins early in the first hour of culture, inactivation did not begin until approximately 40 min after exposing cells to Con A. The 40-min delay in onset of inactivation was divisible into (i) an initial stage needed for cell reaction with Con A and (ii) a subsequent stage which did not need reaction of cells with more Con A molecules. The results indicate that a brief Con A pulse provides sufficient lectin to activate cells to proceed from G0 into the G1 phase of the cell cycle. Once cells have reacted with Con A there are, at least during the first 6 h of culture, no further stages dependent on reaction of cells with more Con A molecules. At high Con A concentrations in the presence of complement, an optimum activation signal is soon countermanded by an optimum inactivation signal. The delay in onset of inactivation is in keeping with previous studies suggesting a need for cell-bound Con A to aggregate cell-surface molecules thus generating receptor sites for complement components.

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Studies on the penetration of mammalian cells by deoxyribonucleoside‐5′‐phosphates

Cited by 6 publications

References 23 publications

Treatment of human tumor cells with ADP or ATP yields arrest of growth in the S phase of the cell cycle

Treatment of human tumor cells with ADP or ATP yields arrest of growth in the S phase of the cell cycle

Increasing transduction efficiency of recombinant murine retrovirus vectors by initiation of endogenous reverse transcription: potential utility for genetic therapies

Lectin pulses as determinants of lymphocyte activation and inactivation during the first six hours of culture: sequential action of concanavalin A and complement cause cell lysis

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